羰基还原酶应用于合成手性2-氯-1-(6-氟-3,4-二氢-2H-1-苯并吡喃-2-基)乙醇  

The application of carbonyl reductase in the synthesis of chiral 2-Chloro-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl) ethanol

作　　者：陈恬 王家洪 王乃星 王福军 宋庆宝[3] 蒲通 CHEN Tian;WANG Jiahong;WANG Naixing;WANG Fujun;SONG Qingbao;PU Tong(Zhejiang Charioteer Pharmaceutical Co.,Ltd.,Zhejiang Xianju 317321,China;Jiangsu Baju Pharmaceutical Co.,Ltd.,Jiangsu Yancheng 224555,China;School of Chemical Engineering ,Zhejiang University of Technology ,Zhejiang Hangzhou 310014,China)

机构地区：[1]浙江车头制药股份有限公司,浙江仙居317321 [2]江苏八巨药业有限公司,江苏盐城224555 [3]浙江工业大学化工学院,浙江杭州310014

出　　处：《南昌大学学报（理科版）》2018年第5期461-466,共6页Journal of Nanchang University(Natural Science)

基　　金：国家国际科技合作专项基金资助项目(2015DFR40360)

摘　　要：2-氯-1-(6-氟-3,4-二氢-2H-1-苯并吡喃-2-基)乙醇(CLA)是合成心血管药物盐酸奈必洛尔的一个前体,有四种构型,即(S,S)-CLA、(S,R)-CLA、(R,S)-CLA和(R,R)-CLA。开发了一种新颖的羰基还原酶专一性地还原2-氯-1-(6-氟-3,4-二氢-2H-1-苯并吡喃-2-基)乙酮(CLK)的酶不对称还原方法制备手性CLA。对反应时间、反应温度和缓冲溶液的pH值进行了条件优化。在25℃～30℃时,在pH为7.5的磷酸盐缓冲溶液中和路易斯酸(氯化镁)存在下,以异丙醇为供氢体,R构型2-氯-1-(6-氟-3,4-二氢-2H-1-苯并吡喃-2-基)乙酮[(R)-CLK]在R构型羰基还原酶/辅酶(β-NAD)体系的作用下,反应24h后专一性地得到(R,R)-CLA产物;(R)-CLK在S构型羰基还原酶/辅酶(β-NAD)体系的作用下,专一性地得到(R,S)-CLA产物;(S)-CLK在(R)-羰基还原酶/辅酶(β-NAD)体系的作用下,专一性地得到(S,R)--CLA产物;和(S)-CLK在(S)-羰基还原酶/辅酶(β-NAD)体系的作用下,专一性地得到(S,S)-CLA产物。所得到的上述四种构型的产物CLA,纯度≥98%,ee(enantiomeric excess,不对映过量)值≥99%,得率≥95%。通过气相色谱与标准样品对照和核磁共振氢谱和碳谱确证了这四种手性产物结构的正确性。方法简单易操作,避免了使用化学还原方法所需要的复杂分离工序,适合于工业化生产。chiral 2-Chloro-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethanol(CLA)is a key precursor for synthesizing cardiovascular drug nebivolol hydrochloride.CLA has four configurations,namely(S,S)-CLA,(S,R)-CLA,(R,S)-CLA and(R,R)-CLA.In this paper,a novel asymmetric reduction of 2-chloro-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethanone(CLK)with carbonyl reductase was investigated.The reaction conditions on reaction time,temperature and pH value of buffer solution were optimized.At 25℃-30℃,in a phosphate buffer solution(pH=7.5)and in the presence of magnesium chloride as a lewis acid and using isopropanol as a hydrogen donor,(R)-2-chloro-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethanone[(R)-CLK]was subjected to(R)-carbonyl reductase/Coenzyme(β-NAD)system for 24 h,and(R,R)-2-Chloro-1-(6-fluoro-3,4-dihydro-2H-1-benzopyran-2-yl)ethanol[(R,R)-CLA]was exclusively produced;the reaction of(R)-CLK with(S)-carbonyl reductase/Coenzyme(β-NAD)system exclusively provided(R,S)-CLA;the reaction of(S)-CLK with(R)-carbonyl reductase/Coenzyme(β-NAD)system exclusively provided(S,R)-CLA;and the reaction of(S)-CLK with(S)-carbonyl reductase/Coenzyme(β-NAD)system exclusively provided(S,S)-CLA.All the four chirally pure CLAs were obtained with purity of≥98%,ee value of≥99%and yield of≥95%,and were all structurally confirmed by comparison with standard samples through gas chromatography and 1H and 13 C NMR spectra.The method is simple,and avoids the complex separation process which was required in the chemical reduction of CLK.Therefore this protocol is applicable in industrial manufacturing.

关 键 词：2-氯-1-(6-氟-3 4-二氢-2H-1-苯并吡喃-2-基)乙醇 2-氯-1-(6-氟-3 4-二氢-2H-1-苯并吡喃-2-基)乙酮 盐酸奈必洛尔 羰基还原酶 不对称还原 

分 类 号：O622.4[理学—有机化学]